1. Additional Considerations for the Care and Use of Amphibians and Reptiles in Research

2. Reptiles -- Special handling considerations - Salmonella - Aggressive/defensive behaviors - biting (all) - clawing (turtles, lizards) - fecal spraying (snakes) Housing Reptiles: Adults and juveniles: moisture, basking sites, density, feeding schedules Eggs: substrate moisture, temp., positioning to avoid membrane damage

3. Euthanasia - Reptiles Intravenous injection of barbituates - problem – barbituates are scheduled drugs (controlled substances) thus difficult to obtain - possible solution – intracoelomic injection of MS-222 (Conroy et al 2009). Inhalation of anesthetics prior to decapitation -- very effective – typical anesthetics employed are isoflurane and halothane Pithing/decapitation Unacceptable method – -- freezing – ice crystals may form prior to death

4. Amphibians -- Special handling considerations: - Salmonella - Toxic dermal secretions (toads, some salamanders) (note – latex gloves may be toxic to amphibians!!) Housing Amphibians: Adults: substrate moisture, hiding substrates, light spectrum, temperature Embryos: aeration, water pH 6 to 7.5 generally ideal, dissolved oxygen Larvae: aeration, pH (as with embryos), feeding (know your species), densities

5. Euthanasia - amphibians Generally similar to reptiles. Also acceptable - dermal application of benzocaine (“Ora-Gel”) followed by decapitation, pithing, or cervical dislocation. Also acceptable – submersion in buffered MS-222 followed by decapitation, pithing, or cervical dislocation

6. GUIDELINES FOR USE OF LIVE AMPHIBIANS AND REPTILES IN FIELD RESEARCH, Compiled by American Society of Ichthyologists and Herpetologists (ASIH), The Herpetologists' League (HL), Society for the Study of Amphibians and Reptiles (SSAR) http://199.245.200.110/pubs/herpcoll.html Numerous other publications dealing with care and use of amphibians and reptiles are linked from the IACUC website http://199.245.200.110/pubs/herpcoll.html